Optimization of temperature and initial pH and kinetic analysis of tartaric acid production by Gluconobacter suboxydans

Response surface methodology was applied for the simultaneous optimization of initial pH and temperature of fermentation for the production of tartaric acid by Gluconobacter suboxydans. A temperature of 32.8v°C and an initial pH of 6.05 was found to be most suitable. Kinetic analysis for the growth of Gluconobacter suboxydans and product formation was done and a logistic model was used to describe the growth of Gluconobacter suboxydans while the Leudeking-Piret model explained the product formation. Parameters of the model were evaluated and tartaric acid formation was found to be non-growth associated.     

Optimization of medium composition for butyric acid production by Clostridium thermobutyricum using response surface methodology

The optimal medium for butyric acid production by Clostridium thermobutyricum in a shake flask culture was studied using statistical experimental design and analysis. The optimal composition of the fermentation medium for maximum butyric acid yield, as determined on the basis of a three-level four-factor Box-Behnken design (BBD), was obtained by response surface methodology (RSM). The high correlation between the predicted and observed values indicated the validity of the model. A maximum butyric acid yield of 12.05 g/l was obtained at K₂HPO₄ 7.2 g/l, 34.9 g/l glucose, 20 g/l yeast extract, and 15 g/l acetate, which compared well to the predicated production of 12.13 g/l.     

响应面设计法优化单葡萄糖醛酸基甘草次酸（GAMG）发酵转化培养基

以甘草酸（dycyfrhizin,GL）为底物,利用产紫青霉（Penicillium purpurogenum Li-3）液态发酵转化单葡萄糖醛酸甘草次酸（GAMG）,采用响应面设计法对初始发酵培养基进行优化。用部分因子分析法研究原始发酵培养基各成分对响应值的显著程度,发现甘草酸（GL）、NaNO3和K2HPO4的质量浓度对发酵产生GAMG的影响显著（P〈0．01）。用中心组合设计确立甘草酸、NaNO3和K2HPO4的适宜质量浓度分别为2．8、3．0和0．8g／L。在优化条件下进行发酵时,GAMG的转化率从75．49％提高到89．11％,比优化前提高了13．62％。     

Optimization of fermentation medium for amidase production by response surface methodology

采用响应面分析方法,对阿萨希丝孢酵母(Trichosporon asahii)ZZB-1产酰胺酶的发酵培养基进行了优化.运用单因子试验筛选出麦芽糖和酵母浸膏为最适碳源、氮源,金属离子Ca2+、Mn2+可提高发酵酰胺酶产量;通过最陡爬坡实验逼近以上4个因子的最大响应区域后,采用Box-Behnken响应面分析法,确定产酰胺酶最佳发酵培养基为麦芽糖18.84 g/L、酵母浸膏9.55 g/L、NaCl 5g/L、KH2 PO41g/L、MgSO4·7H2O 0.2 g/L、FeSO40.001g/L、CaCO370.84 μmol/L、MnSO4 65.39 μmol/L(1％丙烯酸诱导),NH4·H2O调节pH至7.0.培养基优化后酰胺酶产量由初始2554U/L提高到4156 U/L,为原始发酵培养基配方酶活产量的1.63倍.     

响应面法优化产酰胺酶发酵培养基

采用响应面分析方法,对阿萨希丝孢酵母（Trichosporon asahii）ZZB-1产酰胺酶的发酵培养基进行了优化。运用单N子试验筛选出麦芽糖和酵母浸膏为最适碳源、氮源,金属离子Ca^2＋、Mn^2＋可提高发酵酰胺酶产量;通过最陡爬坡实验逼近以上4个因子的最大响应区域后,采用Box—Behnken响应面分析法,确定产酰胺酶最佳发酵培养基为麦芽糖18．84g／L、酵母浸膏9．55g／L、NaC15g／L、KH2PO41g／L、MgSO4·7H2O0．2g／L、FeS040．001g／L、CaC0370．84μmol／L、MnS0465．39肚mo[／L（1％丙烯酸诱导）,NH4·H2O调节pH至7．0。培养基优化后酰胺酶产量由初始2554U／L提高到4156U／L,为原始发酵培养基配方酶活产量的1．63倍。     

Optimization of medium composition for lipase production by Candida rugosa NCIM 3462 using response surface methodology

A sequential optimization approach using statistical design of experiments was employed to enhance the lipase production by Candida rugosa in submerged batch fermentation. Twelve medium components were evaluated initially using the Plackett-Burman 2-level factorial design. The significant variables affecting lipase production were found to be glucose, olive oil, peptone, (NH4)2SO4, and FeCl3.6H2O. Various vegetable oils were tested in the second step, and among them, groundnut oil was found to be the best inducer for lipase production by C. rugosa. The third step was to identify the optimal values of the significant medium components with groundnut oil as the inducer using response surface methodology. The regression equation obtained from the experimental data designed using a central composite design was solved, and analyzing the response surface contour plots, the optimal concentrations of the significant variables were determined. A maximum lipase activity of 5.95 U.mL-1, which is 1.64 times the maximum activity obtained in the Plackett-Burman experimental trials, was observed. The optimum combination of medium constituents contained 19.604 g.L-1 glucose, 13.065 mL.L-1 groundnut oil, 7.473 g.L-1 peptone, 0.962 g.L-1 (NH4)2SO4, 0.0019 g.L-1 FeCl3.6H2O, and other insignificant components at the fixed level. A predictive model of the combined effects of the independent variables using response surface methodology and an artificial neural network was proposed. The unstructured kinetic models, logistic model, and Luedeking-Piret model were used to describe cell mass and lipase production. The parameters of the models were evaluated and the lipase production by C. rugosa was found to be growth associated.     

Box-Behnken响应面法优化司替霉素B产生菌发酵条件

【背景】粗糙链霉菌(Streptomyces scabrisporus) HBERC-53204是本中心自主分离的一株链霉菌,经鉴定,其产生一种活性化合物司替霉素B (steffimycin B,SMB),对多种动植物重要病原菌具有良好生物活性。【目的】提高SMB发酵水平,拓宽放线菌活性天然产物在农牧业领域的研究及应用。【方法】以本实验室筛选出的一株产SMB的粗糙链霉菌HBERC-53204为研究对象,运用单因素试验筛选培养基的主效碳源、氮源、无机盐及各营养成分最适浓度,并基于单因素试验结果,通过Plackett-Burman(PB)试验设计筛选出显著影响因素,再结合最陡爬坡试验、Box-Behnken (BB)响应面法拟合显著因子与产量的非线性方程求解,进一步优化菌株产SMB的最佳发酵培养基配方。【结果】优化后最佳培养基配方为：葡萄糖36.22 g/L,蛋白胨8.00 g/L,酵母粉8.51 g/L,酸水解酪蛋白1.50 g/L,MgSO₄ 0.68 g/L,KNO₃ 1.00 g/L。经摇瓶验证,优化后SMB效价达到477.26 mg/L...     

产S-酰胺酶培养基统计学筛选与响应面优化

利用Design Expert软件中的两水平实验设计和响应面法,对发酵生产S-酰胺酶(可用于拆分2,2-二甲基环丙甲酰胺外消旋体)的培养基进行了优化。采用Plackett-Burman(PB)设计对培养基中相关影响因素的效应进行评价并筛选出了有显著效应的葡萄糖、酵母粉及2,2-二甲基环丙甲酰胺浓度,其他因素对酰胺酶产量的影响不显著。然后用旋转中心组和实验设计及响应面分析确定了主要影响因素的最佳条件,在优化的培养基中,酰胺酶产量达到168 U/L,比优化前的80 U/L提高了110.0%。     

Optimization of medium composition for actinorhodin production by Streptomyces coelicolor A3(2) with response surface methodology

Optimization of the fermentation medium for maximization of actinorhodin production by Streptomyces coelicolor A3(2) was carried out. Response surface methodology (RSM) was applied to optimize the medium constituents. A 2⁴ full-factorial central composite design (CCD) was chosen to explain the combined effects of the four medium constituents, viz. sucrose, glucose, yeast extract (YE) and peptone, and to design a minimum number of experiments. The P-values of the coefficients for linear, quadratic and cross-product effect of sucrose and glucose concentration were <0.0001, suggesting that these were critical variables having the greatest effect on the production of actinorhodin in the complex medium. The optimized medium consisting of 339 g/l sucrose, 1 g/l glucose, 1.95 g/l YE and 2.72 g/l peptone predicted 195 mg/l of actinorhodin which was 32% higher than that of the unoptimized medium. The amounts of glucose, YE and peptone required were also reduced with RSM.     

响应面法优化福鸽霉素发酵培养基

采用Plackett-Burman设计法,对影响纤维堆囊菌So ceMWXAB-125产生福鸽霉素的9个因素进行了筛选。结果表明,影响该菌产生福鸽霉素的主要营养因素为马铃薯淀粉、CaCl2和脱脂奶粉。在此基础上,采用响应面法对其中3个显著因子的最佳水平范围进行研究,利用Design-Expert软件进行二次回归分析得知,马铃薯淀粉、CaCl2和脱脂奶粉的质量浓度分别为8.05、2.72和10.00 g/L时,福鸽霉素的产量从67 mg/L提高到119.98 mg/L。     

Optimization of media composition for Nattokinase production by Bacillus subtilis using response surface methodology

Response surface methodology and central composite rotary design (CCRD) was employed to optimize a fermentation medium for the production of Nattokinase by Bacillus subtilis at pH 7.5. The four variables involved in this study were Glucose, Peptone, CaCl(2), and MgSO(4). The statistical analysis of the results showed that, in the range studied; only peptone had a significant effect on Nattokinase production. The optimized medium containing (%) Glucose: 1, Peptone: 5.5, MgSO(4): 0.2 and CaCl(2): 0.5 resulted in 2-fold increased level of Nattokinase (3194.25U/ml) production compared to initial level (1599.09U/ml) after 10h of fermentation. Nattokinase production was checked with fibrinolytic activity.     

Optimization of the medium composition for production of mycelial biomass and exo-polymer by Grifola frondosa GF9801 using response surface methodology

In this work, a three-level Box-Behnken factorial design was employed combining with response surface methodology (RSM) to optimize the medium composition for the production of the mycelial biomass and exo-polymer in submerged cultures by Grifola frondosa GF9801. A mathematical model was then developed to show the effect of each medium composition and their interactions on the production of mycelial biomass and exo-polymer. The model estimated that, a maximal yield of mycelial biomass (17.61 g/l) could be obtained when the concentrations of glucose, KH2PO4, peptone were set at 45.2 g/l, 2.97 g/l, 6.58 g/l, respectively; while a maximal exo-polymer yield (1.326 g/l) could be achieved when setting concentrations of glucose, KH2PO4, peptone at 58.6 g/l, 4.06 g/l and 3.79 g/l, respectively. These predicted values were also verified by validation experiments. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a significant increase in the yields of mycelial biomass and exo-polymer. Maximum mycelial biomass yield of 22.50 g/l was achieved in a 15-l fermenter using the optimized medium.     

Optimization of medium components for phytase production by E. coli using response surface methodology

The medium for recombinant phytase production by E. coli BL21 was optimized using response surface methodology. A 2³ central composite experimental design was used to study the combined effect of the medium components, tryptone, yeast extract and NaCl. Addition of 2?g/l glucose to the medium greatly influenced the phytase production. The optimization of the medium has increased the phytase production by 1.2 times. The incorporation of 2?g/l glucose significantly enhanced the phytase production by 1.58 times, showing an overall 2.78 fold increase before optimization and other modifications of the medium. The optimized medium with glucose showed a highest phytase activity of 2250?U/l.     

Statistical optimization of the medium composition by response surface methodology to enhance schizophyllan production by Schizophyllum commune

The response surface methodology (RSM) involving central composite design (CCD) was employed to optimize the fermentation medium for the cell growth and schizophllan production by Schizophyllum commune CGMCC 5.113 in submerged culture at pH 6.5 and 26 degrees C. The four variables involved in this study were glucose, yeast extract, ammonium nitrate, and magnesium sulfate. The statistical analysis of the results showed that, in the range studied, glucose and yeast extract had a highly significant effect on schizophyllan production. The optimal medium for schizophyllan production calculated from the regression model of RSM was as follows: glucose, 18 g/l; yeast extract, 0.5 g/l; NH4NO3, 0.48 g/l; and MgSO4, 0.05 g/l, with a predicted maximum schizophyllan production of 11.74 g/l. These predicted values were experimentally validated. The excellent correlation between predicted and measured values justifies the validity of the response model. The results of bioreactor fermentation also show that the optimized medium enhanced schizophyllan production (12.80 g/l) by S. commune in a 5-1 fermenter.     

响应面分析法优化香菇多糖发酵培养基

香菇多糖是香菇中分离出的一种重要的具有生理活性的物质。采用深层发酵技术培养香菇菌丝体生产多糖,可利用农副产品作原料,成本低,周期短,易于大规模生产。采用响应面优化香菇Lentinula edodes135发酵培养基产多糖,首先Plackett-Burman设计对培养基8因素进行筛选,获得影响多糖产量的3个主要因素:葡萄糖,鱼粉和KH2PO4;然后用最陡爬坡路径逼近最大响应区域;最后通过Box-Behnken设计及响应面分析确定了主要影响因素的最佳浓度:葡萄糖2.2533%,鱼粉0.9756%,VB10.003%,NaCl0.8%,MgSO4·7H2O0.1%,FeSO4·7H2O0.04%,KH2PO40.0993%,初始pH值5.5。在优化后的培养基中,粗多糖产量为2.33g/L,实测值与预测值的误差为+3.61%,比初始培养基多糖产量提高1.91倍。     

Optimization of a cultural medium for bacteriocin production by Lactococcus lactis using response surface methodology.

The medium composition for bacteriocin production by Lactococcus lactis ATCC 11454 was optimized using response surface methodology. The selected six factors based on CM medium were sucrose, soybean peptone, yeast extract, KH(2)PO(4), NaCl, and MgSO(4).7H(2)O. Fractional factorial designs (FFD) and the path of steepest ascent were effective in searching for the main factors and approaching the optimum region of the response. By a 2(6-2) FFD, sucrose, soybean peptone, yeast extract, KH(2)PO(4) were found to be significant factors and had positive effects on cell growth, however, only soybean peptone and KH(2)PO(4) were shown to be the two significant factors for bacteriocin production and had negative and positive effects, respectively. The effects of the two main factors on bacteriocin production were further investigated by a central composite design and the optimum composition was found to be 1% sucrose, 0.45% soybean peptone, 1% yeast extract, 2.84% KH(2)PO(4), 0.2% NaCl, and 0.02% MgSO(4) x 7H(2)O. The optimal medium allowed bacteriocin yield to be doubled compared to CM medium.     

响应面法优化嗜热链球菌增殖培养基

目的 为了降低嗜热链球菌工业化生产成本,提高其发酵活菌数,对嗜热链球菌原始培养基的成分进行了响应面优化.方法 利用Design-Expert软件首先采用Plackett-Burman试验设计筛选出了影响嗜热链球菌生长的显著因子,再依据响应面法设计试验,得到嗜热链球菌的优化培养基配比.结果 乳糖、酵母粉和L-半胱氨酸为嗜热链球菌生长的显著性因子;优化后培养基配比由结果分析可知,当活菌数达到最大值时,即:乳糖的浓度为3.6％,酵母粉的浓度为1.74％,L-半胱氨酸的浓度为1.1％,牛肉蛋白胨1％,酪胨1％,KH2PO4 0.20％,Na2 HPO40.20％,叶温-80 0.1％活菌数为最高2.34×109 CFU/mL.结论 采用响应面分析法优化的培养基较经典TPY培养基活菌数提高了近2～3倍.     

Optimization of growth medium for the production of spores from Bacillus thuringiensis using response surface methodology

The effects of cultivation medium compositions including tapioca, fishmeal, CaCO₃ and (NH₄)₂SO₄ for the growth of Bacillus thuringiensis YMB 96-1988 were accessed by using response surface methodology (RSM). The two-level (2^4–1) fractional factorial designs (FFD) which involve two concentrations of each nutrient, and the paths of steepest ascent were effective in searching for the major factors of the bacteria growth. This allows the fitting of a first order linear model to the data. In this study, supplementary CaCO₃ showed a negative effect on the spore production based on the first order regression coefficients derived from SAS programme. Subsequently, a 2³ central composite design (CCD) was used for allocation of treatment combinations. Preliminary studies showed that tapioca and fishmeal is believed to be the major factors for the growth of B. thuringiensis. Estimated optimum compositions for the production of spores by B. thuringiensis are as follows: tapioca, 5.01%; fishmeal 5.86%; (NH₄)₂SO₄ 0.06% and resulted in a maximum spore count of 8.56?×?10⁸/ml was obtained. This value is close to the 8.35?×?10⁸/ml spore density as counted from actual experimental observations.     

Characterization of production of free gluconic acid by Gluconobacter suboxydans adsorbed on ceramic honeycomb monolith

Gluconobacter suboxydans IFO 3290 was immobilized by adsorption on ceramic honeycomb monolith and continuous production of free gluconic acid from glucose was performed in an aerated reactor. The effects of reactor residence time, aeration rate, and glucose concentration were investigated on the gluconic acid yield. Observation of SEM photographs revealed that the cells were adsorbed with a high density not only on the outer surface of the support but also on the inner surface of large pores. From measurement of the number of the adsorbed cells, it was elucidated that the biofilm comprised a monolayer or bilayer of the cells. Maximum specific rate of growth was estimated for the free and adsorbed cells, and the adsorbed cells were found to grow at a fast rate compared with the free cells. In the continuous fermentation performed for one month at the glucose concentration of 100 kg/m(3), reactor residence time of 3.5 h and aeration rate of 900 cm(3)/min, the activity of the adsorbed cells was appreciably stable. The high productivity of 26.3 kg/(m(3)-reactor . h) was attained with the gluconic acid yield of 84.6% and glucose conversion of 94%.